RecombinantRabMAb

Recombinant Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

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Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
  • Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
  • Immunoprecipitation - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
  • Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
  • Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19656] to HIF-2-alpha
  • Suitable for: IP, WB, ICC/IF
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-HIF-2-alpha antibody [EPR19656]
    See all HIF-2-alpha primary antibodies

  • Description

    Rabbit monoclonal [EPR19656] to HIF-2-alpha

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, WB, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Whole cell lysate of HeLa cells treated with 1mM deferoxamine (DFO) mesylate salt for 24h ; Whole cell lysate of HeLa cells treated with 100µM CoCl2 for 24h. ICC/IF: HeLa cells treated with DFO mesylate salt (1mM, 24h).

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab207607 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP
1/30.
WB
1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 96 kDa).
ICC/IF
1/100.
Notes
IP
1/30.
WB
1/1000. Detects a band of approximately 110 kDa (predicted molecular weight: 96 kDa).
ICC/IF
1/100.

Target

  • Function

    Transcription factor involved in the induction of oxygen regulated genes. Binds to core DNA sequence 5'-[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters. Regulates the vascular endothelial growth factor (VEGF) expression and seems to be implicated in the development of blood vessels and the tubular system of lung. May also play a role in the formation of the endothelium that gives rise to the blood brain barrier. Potent activator of the Tie-2 tyrosine kinase expression. Activation seems to require recruitment of transcriptional coactivators such as CREBPB and probably EP300. Interaction with redox regulatory protein APEX seems to activate CTAD.

  • Tissue specificity

    Expressed in most tissues, with highest levels in placenta, lung and heart. Selectively expressed in endothelial cells.

  • Involvement in disease

    Defects in EPAS1 are the cause of erythrocytosis familial type 4 (ECYT4) [MIM:611783]. ECYT4 is an autosomal dominant disorder characterized by increased serum red blood cell mass, elevated hemoglobin concentration and hematocrit, and normal platelet and leukocyte counts.

  • Sequence similarities

    Contains 1 basic helix-loop-helix (bHLH) domain.
    Contains 1 PAC (PAS-associated C-terminal) domain.
    Contains 2 PAS (PER-ARNT-SIM) domains.

  • Post-translational
    modifications

    In normoxia, is probably hydroxylated on Pro-405 and Pro-531 by EGLN1/PHD1, EGLN2/PHD2 and/or EGLN3/PHD3. The hydroxylated prolines promote interaction with VHL, initiating rapid ubiquitination and subsequent proteasomal degradation. Under hypoxia, proline hydroxylation is impaired and ubiquitination is attenuated, resulting in stabilization.
    In normoxia, is hydroxylated on Asn-847 by HIF1AN thus probably abrogating interaction with CREBBP and EP300 and preventing transcriptional activation.
    Phosphorylated on multiple sites in the CTAD.
    The iron and 2-oxoglutarate dependent 3-hydroxylation of asparagine is (S) stereospecific within HIF CTAD domains.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • Basic helix loop helix PAS protein MOP2 antibody
      • Basic-helix-loop-helix-PAS protein MOP2 antibody
      • bHLHe73 antibody
      • Class E basic helix-loop-helix protein 73 antibody
      • ECYT4 antibody
      • Endothelial PAS domain containing protein 1 antibody
      • Endothelial pas domain protein 1 antibody
      • Endothelial PAS domain-containing protein 1 antibody
      • EPAS 1 antibody
      • EPAS-1 antibody
      • EPAS1 antibody
      • EPAS1_HUMAN antibody
      • HIF 1 alpha like factor antibody
      • HIF 2 alpha antibody
      • HIF-1-alpha-like factor antibody
      • HIF-2-alpha antibody
      • HIF2-alpha antibody
      • HIF2A antibody
      • HLF antibody
      • Hypoxia inducible factor 2 alpha antibody
      • Hypoxia inducible factor 2 alpha subunit antibody
      • Hypoxia-inducible factor 2-alpha antibody
      • Member of PAS protein 2 antibody
      • Member of pas superfamily 2 antibody
      • MOP 2 antibody
      • MOP2 antibody
      • PAS domain-containing protein 2 antibody
      • PASD2 antibody
      see all

    Images

    • Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
      Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
      All lanes : Anti-HIF-2-alpha antibody [EPR19656] (ab207607) at 1/1000 dilution

      Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : Whole cell lysate of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 1mM deferoxamine (DFO) mesylate salt for 24h

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 96 kDa
      Observed band size: 110 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The molecular weight observed is consistent with the literature, and the expression level can be induced under hypoxia or treated with hypoxia mimetic chemicals such as DFO (PMID: 25587023, 10934146, 23958427 and 21571835).

    • Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
      Immunocytochemistry/ Immunofluorescence - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, untreated or treated with deferoxamine mesylate salt (1mM, 24h), labeling HIF-2-alpha with ab207607 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing nuclear staining on HeLa cells treated with deferoxamine mesylate salt (1mM, 24h).

      The nuclear counterstain is DAPI (blue).

      Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

    • Immunoprecipitation - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
      Immunoprecipitation - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

      HIF-2-alpha was immunoprecipitated from 0.35 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell treated with 1mM DFO for 24h, cell lysate 10 µg with ab207607 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab207607 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate (10 µg) treated with 1mM DFO for 24h

      Lane 2: ab207607 IP in DFO treated HeLa cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207607 in HeLa cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 75 seconds

       

    • Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
      Western blot - Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
      All lanes : Anti-HIF-2-alpha antibody [EPR19656] (ab207607) at 1/1000 dilution

      Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : Whole cell lysate of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells treated with 100µM CoCl2 for 24h

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 96 kDa
      Observed band size: 110 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The molecular weight observed is consistent with the literature, and the expression level can be induced under hypoxia or treated with hypoxia mimetic chemicals such as CoCl2 (PMID: 21571835, 10934146 and 23958427).

    • Anti-HIF-2-alpha antibody [EPR19656] (ab207607)
      Anti-HIF-2-alpha antibody [EPR19656] (ab207607)

    References (5)

    Publishing research using ab207607? Please let us know so that we can cite the reference in this datasheet.

    ab207607 has been referenced in 5 publications.

    • Magdaleno C  et al. Fibronectin assembly regulates lumen formation in breast acini. J Cell Biochem 122:524-537 (2021). PubMed: 33438770
    • Yao C  et al. SIPA1 Enhances Aerobic Glycolysis Through HIF-2α Pathway to Promote Breast Cancer Metastasis. Front Cell Dev Biol 9:779169 (2021). PubMed: 35096814
    • Magdaleno C  et al. HIFa independent mechanisms in renal carcinoma cells modulate divergent outcomes in fibronectin assembly mediated by hypoxia and CoCl2. Sci Rep 10:18560 (2020). PubMed: 33122751
    • Chen G  et al. Hypoxia induces an endometrial cancer stem-like cell phenotype via HIF-dependent demethylation of SOX2 mRNA. Oncogenesis 9:81 (2020). PubMed: 32913192
    • Yu X  et al. Iron Transport from Ferrous Bisglycinate and Ferrous Sulfate in DMT1-Knockout Human Intestinal Caco-2 Cells. Nutrients 11:N/A (2019). PubMed: 30813537

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